|Publication Type||Journal Article|
|Year of Publication||2019|
|Authors||Adelmann, CH, Wang, T, Sabatini, DM, Lander, ES|
|Journal||Methods Mol Biol|
In this protocol, pooled sgRNA libraries targeting thousands of genes are computationally designed, generated using microarray-based synthesis techniques, and packaged into lentiviral particles. Target cells of interest are transduced with the lentiviral sgRNA pools to generate a collection of knockout mutants-via Cas9-mediated genomic cleavage-and screened for a phenotype of interest. The relative abundance of each mutant in the population can be monitored over time through high-throughput sequencing of the integrated sgRNA expression cassettes. Using this technique, we outline strategies for the identification of cancer driver genes and genes mediating drug response.
|Alternate Journal||Methods Mol. Biol.|